Xu Q, Jürgensmeier J M, Reed J C
Program on Apoptosis and Cell Death Research, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, California 92037, USA.
Methods. 1999 Apr;17(4):292-304. doi: 10.1006/meth.1999.0743.
Bcl-2 family proteins play an evolutionarily conserved role in regulating the life and death of the cell. Certain proapoptotic members of the Bcl-2 family, Bax and Bak, have intrinsic cytotoxic activities in that they not only induce or sensitize mammalian cells to undergo apoptosis but also display a lethal phenotype when ectopically expressed in two yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. Furthermore, the antiapoptotic Bcl-2 and Bcl-XL proteins can protect yeast against Bax-mediated lethality, suggesting that the death-regulatory functions of these Bcl-2 family proteins are well preserved in yeast. These observations provide the opportunity to study the function of Bcl-2 family proteins in genetically tractable yeast and to apply classical yeast genetics and functional cloning approaches to the dissection of programmed cell death pathway regulated by Bcl-2 family proteins. We describe here methods used in our laboratory to express and to study the functions of Bcl-2 family proteins in both the budding yeast S. cerevisiae and the fission yeast S. pombe.
Bcl-2家族蛋白在调节细胞生死过程中发挥着进化上保守的作用。Bcl-2家族的某些促凋亡成员，如Bax和Bak，具有内在的细胞毒性活性，它们不仅能诱导哺乳动物细胞发生凋亡或使其对凋亡敏感，而且当在酿酒酵母和粟酒裂殖酵母这两种酵母中异位表达时，还会表现出致死表型。此外，抗凋亡的Bcl-2和Bcl-XL蛋白可以保护酵母免受Bax介导的致死作用，这表明这些Bcl-2家族蛋白的死亡调节功能在酵母中得到了很好的保留。这些观察结果为在遗传上易于处理的酵母中研究Bcl-2家族蛋白的功能提供了机会，并为剖析由Bcl-2家族蛋白调节的程序性细胞死亡途径应用经典的酵母遗传学和功能克隆方法。我们在此描述了我们实验室用于在芽殖酵母酿酒酵母和裂殖酵母粟酒裂殖酵母中表达和研究Bcl-2家族蛋白功能的方法。
